Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards
Inductively coupled plasma-mass spectrometry (ICP?MS) has been widely used in Life Sciences for the absolute quantification of biomolecules without specific standards, assuming the same response for generic compounds including complex biomolecules. However, contradictory results have been published...
| Authors: | ; ; ; |
|---|---|
| Format: | Article |
| Publication date: | 2020 |
| Country: | España |
| Institution: | Universidad de Cantabria (UC) |
| Repository: | UCrea Repositorio Abierto de la Universidad de Cantabria |
| Language: | English |
| OAI Identifier: | oai:repositorio.unican.es:10902/39484 |
| Online Access: | https://hdl.handle.net/10902/39484 |
| Access Level: | Open access |
| Keyword: | Calibration Hydrophobicity Monomers Peptides and proteins Sulfur |
| _version_ | 1878732141775290368 |
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| author | Cid-Barrio, Laura |
| author2 | Calderón Celis, Francisco Costa-Fernández, José Manuel Ruiz Encinar, Jorge |
| author2_role | author author author |
| author_browse | Calderón Celis, Francisco Cid-Barrio, Laura Costa-Fernández, José Manuel Ruiz Encinar, Jorge |
| author_facet | Cid-Barrio, Laura Calderón Celis, Francisco Costa-Fernández, José Manuel Ruiz Encinar, Jorge |
| author_role | author |
| collection | UCrea Repositorio Abierto de la Universidad de Cantabria |
| description | Inductively coupled plasma-mass spectrometry (ICP?MS) has been widely used in Life Sciences for the absolute quantification of biomolecules without specific standards, assuming the same response for generic compounds including complex biomolecules. However, contradictory results have been published on this regard. We present the first critical statistical comparison of the ICP-MS response factors obtained for 14 different relevant S-containing biomolecules (three peptides, four proteins, one amino acid, two cofactors, three polyethylene glycol (PEG) derivatives, and sulfate standard), covering a wide range of hydrophobicities and molecular sizes. Two regular flow nebulizers and a total consumption nebulizer (TCN) were tested. ICP?MS response factors were determined though calibration curves, and isotope dilution analysis was used to normalize the results. No statistical differences have been found for low-molecular-weight biocompounds, PEGs, and nonhydrophobic peptides using any of the nebulizers tested. Interestingly, while statistical differences were still found negligible (96-104%) for the proteins and hydrophobic peptide using the TCN, significantly lower response factors (87-40%) were obtained using regular flow nebulizers. Such differential behavior seems to be related mostly to hydrophobicity and partially to the molecular weight. Findings were validated using IDA in intact and digested bovine serum albumin solutions using the TCN (98 and 100%, respectively) and the concentric nebulizer (73 and 97%, respectively). Additionally, in the case of a phosphoprotein, results were corroborated using the P trace in parallel to the S trace used along the manuscript. This work seems to suggest that ICP-MS operated with regular nebulizers can offer absolute quantification using generic standards for most biomolecules except proteins and hydrophobic peptides. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_3d655d1a235a00a5a90805ee72ae1df0 |
| institution | Universidad de Cantabria (UC) |
| instname_str | Universidad de Cantabria (UC) |
| language | eng |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:repositorio.unican.es:10902/39484 |
| publishDate | 2020 |
| publishDateSort | 2020 |
| publisher | American Chemical Society |
| publisherStr | American Chemical Society |
| reponame_str | UCrea Repositorio Abierto de la Universidad de Cantabria |
| spelling | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards Cid-Barrio, Laura Calderón Celis, Francisco Costa-Fernández, José Manuel Ruiz Encinar, Jorge Calibration Hydrophobicity Monomers Peptides and proteins Sulfur Inductively coupled plasma-mass spectrometry (ICP?MS) has been widely used in Life Sciences for the absolute quantification of biomolecules without specific standards, assuming the same response for generic compounds including complex biomolecules. However, contradictory results have been published on this regard. We present the first critical statistical comparison of the ICP-MS response factors obtained for 14 different relevant S-containing biomolecules (three peptides, four proteins, one amino acid, two cofactors, three polyethylene glycol (PEG) derivatives, and sulfate standard), covering a wide range of hydrophobicities and molecular sizes. Two regular flow nebulizers and a total consumption nebulizer (TCN) were tested. ICP?MS response factors were determined though calibration curves, and isotope dilution analysis was used to normalize the results. No statistical differences have been found for low-molecular-weight biocompounds, PEGs, and nonhydrophobic peptides using any of the nebulizers tested. Interestingly, while statistical differences were still found negligible (96-104%) for the proteins and hydrophobic peptide using the TCN, significantly lower response factors (87-40%) were obtained using regular flow nebulizers. Such differential behavior seems to be related mostly to hydrophobicity and partially to the molecular weight. Findings were validated using IDA in intact and digested bovine serum albumin solutions using the TCN (98 and 100%, respectively) and the concentric nebulizer (73 and 97%, respectively). Additionally, in the case of a phosphoprotein, results were corroborated using the P trace in parallel to the S trace used along the manuscript. This work seems to suggest that ICP-MS operated with regular nebulizers can offer absolute quantification using generic standards for most biomolecules except proteins and hydrophobic peptides. Financial support from FC-GRUPIN-ID/2018/000166 (Asturias Regional Government) and CTQ2016–79412-P and PID2019- 109698GBI00 (Ministry of Economy and Competitiveness, Spain) is gratefully acknowledged. Authors want to thank also Agilent for the technical support. LCB acknowledges the Ph.D. grant (FPU15/04989) Spanish Ministry of Education. American Chemical Society https://hdl.handle.net/10902/39484 |
| spellingShingle | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards Cid-Barrio, Laura Calibration Hydrophobicity Monomers Peptides and proteins Sulfur |
| title | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards |
| title_full | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards |
| title_fullStr | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards |
| title_full_unstemmed | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards |
| title_short | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards |
| title_sort | Assessment of the potential and limitations of elemental mass spectrometry in life sciences for absolute quantification of biomolecules using generic standards |
| topic | Calibration Hydrophobicity Monomers Peptides and proteins Sulfur |
| topic_facet | Calibration Hydrophobicity Monomers Peptides and proteins Sulfur |
| url | https://hdl.handle.net/10902/39484 |